The research of differentiation of skin-derived precursors into motor neurons induced by retinoic acid combined with Purmorphamine

Abstract

Objective To explore the differentiation of skin-derived precursors (SKPs) into motor neurons (MNs) induced by retinoic acid (RA) combined with Purmorphamine (PM), and provide the evidence for motor neuron transplantation in the treatment of denervated muscle atrophy. Methods SKPs were isolated from the skin of neonatal rats, and cultured and proliferated in vitro. The expression of SKPs markers Nestin and Fibronectin was detected by immunocytochemistry. RA and PM were used as inducers to induce the differentiation of the third generation SKPs to MNs, and the growth factors such as GDNF and BDNF were added to promote the axon elongation and MNs maturity. The expression of MNs marker HB9 and choline acetyltransferase (ChAT) was detected by immunocytochemistry to identify whether the differentiated cells were MNs or not. The positive rate of ChAT and HB9 was calculated. Results The differentiated cells from SKPs induced by RA and PM had the morphology of MNs, and express the MNs specific markers HB9 and ChAT. The positive rate was (72.61±2.25)% for ChAT and (75.16±3.62)% for HB9. Conclusion The use of RA and PM can induce SKPs to differentiate into MNs. Key words: Motor neurons; Cell differentiation; Skin-derived precursors; Retinoic acid